CN104645667A - Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof - Google Patents
Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof Download PDFInfo
- Publication number
- CN104645667A CN104645667A CN201310578325.3A CN201310578325A CN104645667A CN 104645667 A CN104645667 A CN 104645667A CN 201310578325 A CN201310578325 A CN 201310578325A CN 104645667 A CN104645667 A CN 104645667A
- Authority
- CN
- China
- Prior art keywords
- port
- chromatogram
- triple valve
- way valve
- pipeline
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to an expanded bed chromatography and counter-current chromatography on-line combination method, an application and an apparatus thereof. The method comprises the following steps: 1)placing a natural product in an extraction apparatus, adding an extraction reagent for extracting; 2)performing recycle extraction; 3)after extracting, eluting; 4)conveying an eluate to an interface through a pump, completing the treatment and collection of a first part sample; 5)pumping the prepared stationary phase in a counter current chromatographic column by the pump, after the stationary phase is fully filled, adjusting the rotating speed of a host to a set value, and pumping a mobile phase at a constant flow velocity; 6)after the stationary phase and the mobile phase reach the balance in the chromatographic column, switching a sampling valve at an interface position, injecting a sample solution into the interface; 7)after the sample solution is treated, switching the sampling valve, injecting the sample into the counter current chromatograph for separating; and 8)adjusting a detector, according to the spectra of the detector, using an automatic fraction collector for collecting target fractions. The method has the advantages of short extraction time, high automation degree and little object component loss.
Description
[technical field]
The present invention relates to chromatogram arrangement technical field, specifically, is a kind of expansion bed chromatogram and adverse current chromatogram on-line coupling method, application and device.
[background technology]
The chemical composition that natural products comprises is very complicated, and the active ingredient wherein playing curative effect effect is of a great variety, comprises the natural components such as albumen, polysaccharide, amino acid, alkaloid, volatile oil, flavonoids, tannin, terpene, steroidal compounds.Extraction and isolation active ingredient and active component can reduce the toxicity of crude drug, improve its curative effect, have great significance for the development of promotion natural products.One of key factor of restriction natural products pharmacy industry is the Extraction and separation to active ingredient, traditional extraction separation method, and ubiquity that effective component extraction rate is low, energy consumption is high, operation is many and the shortcoming such as the production cycle is long.Along with the acceleration of China's modernization of Chinese medicine process, the quality standard of Chinese medicine Related product improves constantly, particularly the carrying out of finger-print and multi-target ingredient quantitative study work, to the quantity of Chinese medicine reference substance and the demand of kind also increasing.On the other hand, the monomeric compound in Chinese medicine is also the important sources of Western medicine and lead compound.In the development process of these medicines, medicine efficacy screening needs a certain amount of monomeric compound, and the modification of lead compound more needs a certain amount of monomeric compound.From natural products, extracting directly monomer component can reduce costs greatly.Along with the development of modern science and technology, in medicament research and development, reducing impurity to reach, improving drug effect, reduce costs these targets, the high-new engineering in some modern times is constantly produced to medicine by reference.
Expanded Bed Adsorption (Expanded Bed Adsorption, EBA) be the novel separating and purifying technology that last century, the nineties grew up, integrate the novel integrated operating process of clarification, concentrated and preliminary purification, the unit operations steps such as centrifugal or filtration can be reduced, reduce the cost of separation and purification, simplify purifying process, be one and apply wide isolation technics.Current expansion bed is mainly used in the separation and purification of biological sample, less be applied to Chinese medicine from.Integrated separation focuses in expansion bed separation process by Separation of Solid and Liquid and Liquid liquid Separation, realizes the easy of operation and the high efficiency of process.
Expansion bed, as a kind of fluidization of particularization, operates under low flow velocity condition, can make the classification that bed reaches stable, therefore back-mixing is less, close to fixed bed.Expanded Bed Adsorption agent has certain particle diameter and Density Distribution, and the absorbent particles that particle diameter is large or density is large is in operation distributed in bottom bed, and the distribution of particles that particle diameter is little or density is little is on bed top.Control suitable operating condition, the back-mixing motion of particle in bed body just can be made to be restricted, thus make adsorbent reach the requirement of layering.Individual particle only moves in a circle among a small circle, and liquid phase flowing is piston flow.Expansion bed bed body after layering has the voidage higher than fixed bed, and this can make solid particle pass a body, and this is also the reason that expansion bed can process containing solid particle extract.
High-speed countercurrent chromatography (High-Speed Countercurrent Chromatography, HSCCC) be the current more novel liquid without solid carrier-liquid distribution technique in the world, developed by US National health research institute professor Ito at first, through the research and development in more than ten years, CCC is further developed into high speed adverse current chromatogram.Owing to which overcoming the dead suction-operated of conventional solid carrier to sample, and be very applicable to Natural products research work to relevant analysis, isolation technics coupling.
High speed adverse current chromatogram has significantly different compared with traditional chromatography, main in the following aspects: (1) due to high speed adverse current chromatogram be continuous print liquid liquid partition chromatography, without the need to solid support, avoid the problem such as irreversibility absorption, decomposition of sample in separation process, be applicable to the separation of active skull cap components.(2) because liquid is as mobile phase and Stationary liquid, the sample be trapped in post can be reclaimed completely by multiple type of elution, can complete separation and purification and preparation simultaneously, is suitable for preparation property and is separated.(3) high speed adverse current chromatogram by the change to solvent system, can be separated the compound of opposed polarity scope, meets the complexity of natural products, multifarious feature especially.(4) compared with HPLC, HSCCC sample size is comparatively large, and can reach several grams at most, be hundreds of times of HPLC.Compared with normal pressure, low-pressure chromatography.The separating power of HSCCC is strong, and namely some sample obtains 1 even multiple compound through first separation, and disengaging time is short, and a few hours can complete, and purity is many more than 98%.(5) can adopt multiple type of elution, as equal proportion wash-out, gradient elution, or two-way type of elution is effectively separated sample.The fields such as biological medicine, agricultural food product, particularly Natural Medicine Chemistry are widely used in based on above feature high speed adverse current chromatogram.
On-line coupling technology is the technology two or more method being combined use by interface, they are learnt from other's strong points to offset one's weaknesses mutually, synergy, not only can improve sensitivity, the degree of accuracy of method, also may obtain function not available when two kinds of methods are used alone.Therefore, on-line coupling technology is one of Disciplinary Frontiers of analytical chemistry research always.Wherein, hyphenated techniques chromatography is with the fastest developing speed.Chromatogram, as the very effective separation method of one, usually with other qualitative method couplings, realizes the analysis to complex mixture.On the other hand, a kind of chromatogram also can with another chromatographic process coupling (Two way chromatograms), to play different clastotype chromatogram distinctive separating power separately.
Although hyphenated techniques chromatography has developed for many years, it has been still one of the study hotspot in analytical chemistry field.Hyphenated techniques chromatography is just towards the future development such as multidimensional, small scale, and the Method and Technology related to is also increasing, as the separation methods such as gas-chromatography, high performance liquid chromatography, supercritical fluid chromatography and ultraviolet, mass spectrum, infrared, the analytical method coupling such as nuclear magnetic resonance, near-infrared.Multi-dimensional chromatograph isolation technics sample is injected multiple separation systems of orthogonal thereto distribution, and in sample, each component is that initial point launches on the detaching direction of multidimensional with sample delivery point.Its advantage is that separating power is strong, and peak capacity is high, eliminates complicated pre-treatment step, can be used for the separation and analysis of extremely complex system.Following GC-MS can become the powerful measure solving conventional analysis problem.
By expansion bed and adverse current chromatograph joint used, the advantage of the two can be given full play to.Expansion bed chromatogram is used for extraction and the first dimension initial gross separation of natural products, and obtained component is transferred to the second dimension high speed adverse current chromatogram by interface and is further purified, and obtains target compound.As can be seen here, expansion bed and high speed adverse current chromatograph joint used system can realize one step from natural products and obtain monomeric compound, be a kind of fast, the chromatogram on-line coupling pattern that efficient, loss is little.
[summary of the invention]
The object of the invention is to overcome the deficiencies in the prior art, a kind of expansion bed chromatogram and adverse current chromatogram on-line coupling method, application and device are provided; Multiple operating procedures such as natural products material component can extract by the present invention, concentrate, purifying are integrated in a step and complete, and greatly can shorten the operating time, reduce sample loss, improve yield, reduce production cost.By the operation of a unit, the method and apparatus of the extraction and isolation natural products effective monomer component of effective monomer component just directly can be isolated from natural products material.
The object of the invention is to be achieved through the following technical solutions:
A device for expansion bed chromatogram and adverse current chromatogram on-line coupling, it comprises extraction element, it is characterized in that, extraction element connects the first triple valve by pipeline; First triple valve connects the first fluid reservoir and peristaltic pump respectively by pipeline; Peristaltic pump connects the second triple valve by pipeline; Second triple valve connects the 3rd triple valve and the 4th triple valve by pipeline; Wherein the 4th triple valve connects expansion bed chromatographic column top and extraction element by pipeline; 3rd triple valve connects expansion bed column bottom and the first detector by pipeline; Detector connects the first port I of six-way valve by pipeline; Second port II of six-way valve, the five-port V of six-way valve connects connecting interface by pipeline; 3rd port III of six-way valve connects constant flow pump by pipeline; Constant flow pump connects the second fluid reservoir by pipeline; 4th port IV of six-way valve connects counter-current chromatograph by pipeline; 6th port VI of six-way valve connects the 3rd fluid reservoir by pipeline; Counter-current chromatograph connects the second detector by pipeline; Second detector connects automatic fraction collector by pipeline.
Described extraction element is the extractor of band heating jacket, ultrasonic extraction device, Microwave Extraction device, percolating extractor, the one in bionic extraction device or semi-bionic extraction method device.
Described connecting interface is quantitative loop interface, the one in trapping column interface or vacuum based solvent evaporation interface.
Described detector is UV-detector, the one in EISD or fluorescence detector.
A kind of expansion bed chromatogram and high speed adverse current chromatogram on-line coupling device step in natural products prepare the application in pure compound.
Expansion bed chromatogram and high speed adverse current chromatogram on-line coupling device step in natural products prepare an application process for pure compound, and it comprises the following steps:
(1) natural products is placed in extraction element, adds extraction reagent, extract;
Described natural products is animal, and plant and microorganism etc. are multiple containing bioactivator;
Described extraction reagent is methanol solution, ethanolic solution, inorganic salt solution, dilute acid soln or dilute alkaline soln;
(2) while extracting, with peristaltic pump, extract in extraction element is passed through the first triple valve, second triple valve, 3rd triple valve is from bottom to top delivered in expansion bed chromatographic column, after the adsorbent media adsorbs in expansion bed chromatographic column, the remaining liquid of absorption is flow back in extraction element by the 4th triple valve and continues to participate in extracting, and is formed and extracts the circulation of medicinal material;
Filler used in described expansion bed chromatographic column is macroporous absorbent resin, ion exchange resin, cellular glass, and the expansion bed such as glucan-silica gel commonly use adsorbing base;
(3) extract complete, close extraction element, by peristaltic pump by the eluting solvent in the first fluid reservoir by the first triple valve, the second triple valve, the 4th triple valve is from top to bottom delivered to expansion bed chromatographic column from expansion bed chromatographic column top, carries out wash-out;
(4) by peristaltic pump by eluent via the first fluid reservoir, first triple valve, second triple valve, 3rd triple valve, first detector, the first port I of six-way valve, the second port II of six-way valve, the five-port V of six-way valve carries connecting interface, completes process and the collection of Part I sample;
(5) with constant flow pump, the Stationary liquid in the second fluid reservoir is pumped in counter-current chromatograph, to be fixed be full of chromatographic column mutually after, counter-current chromatograph engine speed is adjusted to setting value, pumps into the mobile phase in the second fluid reservoir with constant flow rate;
(7) after two-phase reaches balance in counter-current chromatograph, switch six-way valve, together with the first port I of six-way valve and the 6th port VI of six-way valve, the five-port V of six-way valve and the 4th port IV of six-way valve, 3rd port III of six-way valve and the second port II of six-way valve, injects counter-current chromatograph with constant flow pump by the sample solution in connecting interface and is separated;
(8) regulate the second detector, according to detector spectrogram, use automatic fraction collector to collect target component;
Described target component refers to the active ingredient of the natural products of preextraction or separation;
Compared with prior art, good effect of the present invention is:
(1) the present invention's one step from natural products prepares the expansion bed chromatogram of pure compound and high speed adverse current chromatogram on-line coupling method and device, gathered in the past centrifugal in natural product extraction and adverse current chromatogram sample pretreatment process, filter, multiple step such as concentrated, redissolution in an operating unit, have the advantages that the time is short, efficiency is high;
(2) the present invention and device adopt totally closed operation environment, decrease the contact of external environment, can avoid pollution and the loss of sample;
(3) because extraction and isolation of the present invention is consuming time short, the time of contact of sample and thermal source can be reduced, the loss that sample degradation causes can be prevented.
[accompanying drawing explanation]
Fig. 1 structural representation of the present invention;
Being labeled as in accompanying drawing: 1 extraction element, 2 first triple valves, 3 first fluid reservoirs, 4 peristaltic pumps, 5 second triple valves, 6 the 3rd triple valves, 7 the 4th triple valves, 8 expansion bed chromatographic columns, 9 first detectors, 10 six-way valves, 11 constant flow pumps, 12 second fluid reservoirs, 13 the 3rd fluid reservoirs, 14 connecting interfaces, 15 counter-current chromatographs, 16 second detectors, 17 automatic fraction collectors; I first port, II second port, III the 3rd port, IV the 4th ports, V five-port, VI the 6th ports.
[detailed description of the invention]
The detailed description of the invention of a kind of expansion bed chromatogram of the present invention and adverse current chromatogram on-line coupling method, application and device is below provided.
Embodiment 1
Refer to accompanying drawing 1, the device of a kind of expansion bed chromatogram and adverse current chromatogram on-line coupling, it comprises extraction element, and extraction element 1 connects the first triple valve 2 by pipeline; First triple valve 2 connects the first fluid reservoir 3 and peristaltic pump 4 respectively by pipeline; Peristaltic pump 4 connects the second triple valve 5 by pipeline; Second triple valve 5 connects the 3rd triple valve 6 and the 4th triple valve 7 by pipeline; Wherein the 4th triple valve 7 connects expansion bed chromatographic column 8 top and extraction element 1 by pipeline; 3rd triple valve 6 connects bottom expansion bed chromatographic column 8 and the first detector 9 by pipeline; Detector 9 connects six-way valve 10 first port I by pipeline; Six-way valve 10 second port II, five-port V connect connecting interface 14 by pipeline; Six-way valve the 3rd port III connects constant flow pump 11 by pipeline; Described constant flow pump 11 connects the second fluid reservoir 12 by pipeline; Described six-way valve the 4th port IV connects counter-current chromatograph 15 by pipeline; Six-way valve the 6th port VI connects the 3rd fluid reservoir 13 by pipeline; Counter-current chromatograph 15 connects the second detector 16 by pipeline; Second detector 16 connects automatic fraction collector 17 by pipeline.
Expansion bed chromatogram and high speed adverse current chromatogram on-line coupling device step in natural products prepare an application process for pure compound, and it comprises the following steps:
(1) natural products is placed in extraction element 1, adds extraction reagent, extract; (2) while extracting, with peristaltic pump 4 by the extract in extraction element 1 through the first triple valve 2, second triple valve 5, the 3rd triple valve 6, from bottom to top be delivered in expansion bed chromatographic column 8, the remaining liquid of absorption flows back in extraction element 1 through the 4th triple valve 7 and continues to participate in extracting, and is formed and extracts the circulation of medicinal material; (3) extract complete, close extraction element 1, switch the first triple valve 2, second triple valve 5, the 3rd triple valve 6, the 4th triple valve 7, by peristaltic pump 4, eluting solvent in first fluid reservoir 3 is from top to bottom delivered to expansion bed chromatographic column 8 from expansion bed chromatographic column 8 top, carries out wash-out; (4) by peristaltic pump 4, eluent in first fluid reservoir 3 is delivered to connecting interface 14 through the first triple valve 2, second triple valve 5, the 3rd triple valve 6, detector 9, six-way valve 10 first port I, the second port II, complete process and the collection of Part I sample, remaining liquid flow to the 3rd fluid reservoir 13 through six-way valve 10 five-port V, the 6th port VI; (5) with constant flow pump 11, adverse current chromatogram Stationary liquid off-the-shelf in second fluid reservoir 12 is pumped in counter-current chromatograph 15 through six-way valve 10 the 3rd port III, the 4th port IV, after adverse current chromatogram Stationary liquid is full of, counter-current chromatograph 15 engine speed is adjusted to setting value, pumps into the adverse current chromatogram mobile phase in fluid reservoir 12 with constant flow rate; (6) after two-phase reaches balance in counter-current chromatograph 15, switch six-way valve 10, with three port III, the second port II of the adverse current chromatogram mobile phase in fluid reservoir 12 through six-way valve 10, the sample in connecting interface is injected counter-current chromatograph 15 be separated through the five-port V of six-way valve 10, the 4th port IV; (8) regulate the second detector 16, according to the second detector 16 spectrogram, use automatic fraction collector 17 to collect target fraction.
By the extraction and isolation of apparatus of the present invention to effective component in red sage, describe the present invention's one step from natural products in detail and prepare the expansion bed chromatogram of pure compound and the method for high speed adverse current chromatogram on-line coupling, the inventive method comprises the following steps:
(1) salvia piece 37g is placed in extraction element 1, adds extract (the present embodiment adopts 250mL water), start extraction element 1, the red sage root in extraction element 1 is extracted;
(2) while extraction, with peristaltic pump 4 by the extract in extraction element 1 through the first triple valve 2, second triple valve 5, the 3rd triple valve 6, from bottom to top be delivered in expansion bed chromatographic column 8, extract is by after adsorbent media adsorbs, the remaining liquid of absorption flows back in extraction element 1 through the 4th triple valve 7 and continues to participate in extracting, and is formed and extracts the circulation of medicinal material;
(3) extract complete, close extraction element 1, switch the first triple valve 2, second triple valve 5, the 3rd triple valve 6, the 4th triple valve 7, by peristaltic pump 4, eluting solvent in first fluid reservoir 3 is from top to bottom delivered to expansion bed chromatographic column 8 from expansion bed chromatographic column 8 top, carry out wash-out with 4 times of column volumes, eluent flow in the 3rd fluid reservoir 13 store through the 3rd triple valve 6, detector 9, six-way valve 10 first port I, the second port II, connecting interface 14, six-way valve 10 five-port V, the 6th port VI;
(4) by peristaltic pump 4, eluent in first fluid reservoir 3 is conducted through detector 9 through the first triple valve 2, second triple valve 5, the 3rd triple valve 6, after effective component in red sage in the eluent be detected is higher than preset value, high concentration eluent is full of quantitative loop interface 14, switch six-way valve 10, make the first port I and second port II of six-way valve 10,3rd port III and the 4th port IV, five-port V is communicated with the 6th port VI, treat that sample solution is full of quantitative loop interface 14, complete process and the collection of Part I sample, remaining liquid flow to the 3rd fluid reservoir 13;
(5) adverse current chromatogram dicyandiamide solution is prepared, using the upper and lower Stationary liquid as adverse current chromatogram and mobile phase; (6) with constant flow pump 11, Stationary liquid is pumped in counter-current chromatograph 15 through the 3rd port III of six-way valve 10 and the 4th port IV from the second fluid reservoir 12, to be fixed be full of counter-current chromatograph 15 mutually after, counter-current chromatograph 15 engine speed is adjusted to setting value, then with constant flow rate, mobile phase is pumped into from the second fluid reservoir 12;
(7) after two-phase reaches balance in counter-current chromatograph 15, switch six-way valve 10, by constant flow pump by mobile phase, through the 3rd port III, the second port II of six-way valve 10 by the sample solution in connecting interface 14 by five-port V, the 4th port IV of six-way valve 10, inject adverse current chromatogram 15 and be separated;
(8) regulate the second detector 16, according to detector spectrogram, use automatic fraction collector 17 to collect target component.
Adopt the inventive method and device, to the extraction purification result of the red sage root, compared with off-line extraction separation method, its Comparative result is as follows:
The situation that table 1 the present invention is separated 37g effective component in red sage with off-line method in a step contrasts
As can be seen from table 1 comparing result, this method and device have environmental protection, quick, step simplifies, efficiency is high, the advantages such as principal product (tanshin polyphenolic acid B) purity is good.
Embodiment 3
By the extraction and isolation of apparatus of the present invention to coptis active ingredient, describe the present invention's one step from natural products in detail and prepare the expansion bed chromatogram of pure compound and the method for high speed adverse current chromatogram on-line coupling, the inventive method comprises the following steps:
(1) coptis medicine materical crude slice 10g is placed in extraction element 1, adds extract (the present embodiment adopts 150mI water), start extraction element 1, the coptis in extraction element 1 is extracted;
(2) while extraction, with peristaltic pump 4 by the extract in extraction element 1 through the first triple valve 2, second triple valve 5, the 3rd triple valve 6, from bottom to top be delivered in expansion bed chromatographic column 8, extract is by after adsorbent media adsorbs, the remaining liquid of absorption flows back in extraction element 1 through the 4th triple valve 7 and continues to participate in extracting, and is formed and extracts the circulation of medicinal material;
(3) extract complete, close extraction element 1, switch the first triple valve 2, second triple valve 5, the 3rd triple valve 6, the 4th triple valve 7, by peristaltic pump 4, eluting solvent in first fluid reservoir 3 is from top to bottom delivered to expansion bed chromatographic column 8 from expansion bed chromatographic column 8 top, carry out wash-out with 4 times of column volumes, eluent flow in the 3rd fluid reservoir 13 store through the 3rd triple valve 6, detector 9, six-way valve 10 first port I, the second port II, connecting interface 14, six-way valve 10 five-port V, the 6th port VI;
(4) by peristaltic pump 4, eluent in first fluid reservoir 3 is conducted through detector 9 through the first triple valve 2, second triple valve 5, the 3rd triple valve 6, after coptis active ingredient in the eluent be detected is higher than preset value, high concentration eluent is full of quantitative loop interface 14, switch six-way valve 10, make the first port I and the second port II of six-way valve 10,3rd port III and the 4th port IV, five-port V is communicated with the 6th port VI, treat that sample solution is full of quantitative loop interface 14, complete process and the collection of Part I sample, remaining liquid flow to the 3rd fluid reservoir 13;
(5) adverse current chromatogram dicyandiamide solution is prepared, using the upper and lower Stationary liquid as adverse current chromatogram and mobile phase;
(6) with constant flow pump 11, Stationary liquid is pumped in counter-current chromatograph 15 through the 3rd port III of six-way valve 10 and the 4th port IV from the second fluid reservoir 12, to be fixed be full of counter-current chromatograph 15 mutually after, counter-current chromatograph 15 engine speed is adjusted to setting value, then with constant flow rate, mobile phase is pumped into from the second fluid reservoir 12;
(7) after two-phase reaches balance in counter-current chromatograph 15, switch six-way valve 10, by constant flow pump by mobile phase, through the 3rd port III, the second port II of six-way valve 10 by the sample solution in connecting interface 14 by five-port V, the 4th port IV of six-way valve 10, inject adverse current chromatogram 15 and be separated;
(8) regulate the second detector 16, according to detector spectrogram, use automatic fraction collector 17 to collect target component.
Adopt the inventive method and device, to the extraction purification result of the coptis, compared with off-line extraction separation method, its Comparative result is as follows:
The situation that table 2 the present invention is separated 10g coptis active ingredient with off-line method in a step contrasts
As can be seen from table 2 comparing result, this method and device have environmental protection, quick, step simplifies, efficiency is high, the advantages such as product purity is good.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, without departing from the inventive concept of the premise; can also make some improvements and modifications, these improvements and modifications also should be considered within the scope of protection of the present invention.
Claims (10)
1. a device for expansion bed chromatogram and adverse current chromatogram on-line coupling, it comprises extraction element, it is characterized in that, extraction element connects the first triple valve by pipeline; First triple valve connects the first fluid reservoir and peristaltic pump respectively by pipeline; Peristaltic pump connects the second triple valve by pipeline; Second triple valve connects the 3rd triple valve and the 4th triple valve by pipeline; Wherein the 4th triple valve connects expansion bed chromatographic column top and extraction element by pipeline; 3rd triple valve connects expansion bed column bottom and the first detector by pipeline; Detector connects the first port I of six-way valve by pipeline; Second port II of six-way valve, the five-port V of six-way valve connects connecting interface by pipeline; 3rd port III of six-way valve connects constant flow pump by pipeline; Constant flow pump connects the second fluid reservoir by pipeline; 4th port IV of six-way valve connects counter-current chromatograph by pipeline; 6th port VI of six-way valve connects the 3rd fluid reservoir by pipeline; Counter-current chromatograph connects the second detector by pipeline; Second detector connects automatic fraction collector by pipeline.
2. the device of a kind of expansion bed chromatogram as claimed in claim 1 and adverse current chromatogram on-line coupling, is characterized in that, described extraction element is the extractor of band heating jacket, ultrasonic extraction device, Microwave Extraction device, percolating extractor, the one in bionic extraction device or semi-bionic extraction method device.
3. the device of a kind of expansion bed chromatogram as claimed in claim 1 and adverse current chromatogram on-line coupling, is characterized in that, described connecting interface is quantitative loop interface, the one in trapping column interface or vacuum based solvent evaporation interface.
4. the device of a kind of expansion bed chromatogram as claimed in claim 1 and adverse current chromatogram on-line coupling, is characterized in that, described detector is UV-detector, the one in EISD or fluorescence detector.
5. an expansion bed chromatogram and high speed adverse current chromatogram on-line coupling device step in natural products prepare the application in pure compound.
6. expansion bed chromatogram and high speed adverse current chromatogram on-line coupling device step in natural products prepare an application process for pure compound, and it is characterized in that, it comprises the following steps:
(1) natural products is placed in extraction element, adds extraction reagent, extract;
(2) while extracting, with peristaltic pump, extract in extraction element is passed through the first triple valve, the second triple valve, 3rd triple valve is from bottom to top delivered in expansion bed chromatographic column, after the adsorbent media adsorbs in expansion bed chromatographic column, the remaining liquid of absorption is flow back in extraction element by the 4th triple valve and continues to participate in extracting, and is formed and extracts the circulation of medicinal material;
(3) extract complete, close extraction element, by peristaltic pump by the eluting solvent in the first fluid reservoir by the first triple valve, the second triple valve, the 4th triple valve is from top to bottom delivered to expansion bed chromatographic column from expansion bed chromatographic column top, carries out wash-out;
(4) by peristaltic pump by eluent via the first fluid reservoir, first triple valve, second triple valve, the 3rd triple valve, first detector, first port I of six-way valve, second port II of six-way valve, the five-port V of six-way valve carries connecting interface, completes process and the collection of Part I sample;
(5) with constant flow pump, the Stationary liquid in the second fluid reservoir is pumped in counter-current chromatograph, to be fixed be full of chromatographic column mutually after, counter-current chromatograph engine speed is adjusted to setting value, pumps into the mobile phase in the second fluid reservoir with constant flow rate;
(7) after two-phase reaches balance in counter-current chromatograph, switch six-way valve, together with the first port I of six-way valve and the 6th port VI of six-way valve, the five-port V of six-way valve and the 4th port IV of six-way valve, 3rd port III of six-way valve and the second port II of six-way valve, injects counter-current chromatograph with constant flow pump by the sample solution in connecting interface and is separated;
(8) regulate the second detector, according to detector spectrogram, use automatic fraction collector to collect target component.
7. a kind of expansion bed chromatogram as claimed in claim 6 and high speed adverse current chromatogram on-line coupling device step in natural products prepare the application process of pure compound, it is characterized in that, in described step (1), described natural products is that animal, plant and microorganism etc. are multiple containing bioactivator.
8. a kind of expansion bed chromatogram as claimed in claim 6 and high speed adverse current chromatogram on-line coupling device step in natural products prepare the application process of pure compound, it is characterized in that, in described step (1), described extraction reagent is methanol solution, ethanolic solution, inorganic salt solution, dilute acid soln or dilute alkaline soln.
9. a kind of expansion bed chromatogram as claimed in claim 6 and high speed adverse current chromatogram on-line coupling device step in natural products prepare the application process of pure compound, it is characterized in that, in described step (2), filler used in described expansion bed chromatographic column is macroporous absorbent resin, ion exchange resin, cellular glass, the expansion bed such as glucan-silica gel commonly use adsorbing base.
10. a kind of expansion bed chromatogram as claimed in claim 6 and high speed adverse current chromatogram on-line coupling device step in natural products prepare the application process of pure compound, it is characterized in that, in described step (8), described target component refers to the active ingredient of the natural products of preextraction or separation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310578325.3A CN104645667B (en) | 2013-11-18 | 2013-11-18 | Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310578325.3A CN104645667B (en) | 2013-11-18 | 2013-11-18 | Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104645667A true CN104645667A (en) | 2015-05-27 |
| CN104645667B CN104645667B (en) | 2017-02-08 |
Family
ID=53237613
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310578325.3A Expired - Fee Related CN104645667B (en) | 2013-11-18 | 2013-11-18 | Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104645667B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108254475A (en) * | 2018-03-09 | 2018-07-06 | 山东省分析测试中心 | A kind of high-speed counter-current chromatograph and method of work for complex sample multi-cycle separation |
| CN113402635A (en) * | 2021-06-18 | 2021-09-17 | 汕头大学 | Method for separating and purifying heparin by expanded bed countercurrent chromatography |
| CN113504330A (en) * | 2021-06-18 | 2021-10-15 | 安莱博医药(苏州)有限公司 | Process and device for purifying oligomeric dimethicone |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4042499A (en) * | 1976-08-02 | 1977-08-16 | The Dow Chemical Company | Liquid adsorption chromatographic apparatus and method |
| US5071547A (en) * | 1990-03-23 | 1991-12-10 | Separations Technology, Inc. | Column chromatographic column apparatus with switching capability |
| CN101158668A (en) * | 2007-10-19 | 2008-04-09 | 中山大学 | Microwave auxiliary extract-high speed adverse current chromatograph joint used method and device thereof |
| CN101386815A (en) * | 2007-09-11 | 2009-03-18 | 中国科学院过程工程研究所 | Coupling apparatus and technique for fermentation and separation of succinic acid by expanded bed adsorption and in situ extraction |
| CN101637665A (en) * | 2009-01-09 | 2010-02-03 | 中山大学 | Method for combined use of accelerated solvent extraction and high speed countercurrent chromatogram and device thereof |
-
2013
- 2013-11-18 CN CN201310578325.3A patent/CN104645667B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4042499A (en) * | 1976-08-02 | 1977-08-16 | The Dow Chemical Company | Liquid adsorption chromatographic apparatus and method |
| US5071547A (en) * | 1990-03-23 | 1991-12-10 | Separations Technology, Inc. | Column chromatographic column apparatus with switching capability |
| CN101386815A (en) * | 2007-09-11 | 2009-03-18 | 中国科学院过程工程研究所 | Coupling apparatus and technique for fermentation and separation of succinic acid by expanded bed adsorption and in situ extraction |
| CN101158668A (en) * | 2007-10-19 | 2008-04-09 | 中山大学 | Microwave auxiliary extract-high speed adverse current chromatograph joint used method and device thereof |
| CN101637665A (en) * | 2009-01-09 | 2010-02-03 | 中山大学 | Method for combined use of accelerated solvent extraction and high speed countercurrent chromatogram and device thereof |
Non-Patent Citations (1)
| Title |
|---|
| 程屹俊: "人参不同层次化学物质组的绿色化学制备方法研究", 《中国博士学位论文全文数据库工程科技I辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108254475A (en) * | 2018-03-09 | 2018-07-06 | 山东省分析测试中心 | A kind of high-speed counter-current chromatograph and method of work for complex sample multi-cycle separation |
| CN113402635A (en) * | 2021-06-18 | 2021-09-17 | 汕头大学 | Method for separating and purifying heparin by expanded bed countercurrent chromatography |
| CN113504330A (en) * | 2021-06-18 | 2021-10-15 | 安莱博医药(苏州)有限公司 | Process and device for purifying oligomeric dimethicone |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104645667B (en) | 2017-02-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104713973B (en) | There is two-dimentional preparative hplc instrument system and the application thereof of on-line preconcentration function | |
| CN102584918B (en) | Method for preparing high-purity baicalin | |
| CN103408602B (en) | A kind of method being separated preparation four kinds of glycoside chemical reference substances from ZANGYINCHEN | |
| CN100439384C (en) | Method for separating preparing anthocyan monomer from mulberry | |
| CN102580352B (en) | Separation method of natural product | |
| CN104415573A (en) | Method for classifying and preparing tertiary amine alkaloid and quaternary ammonium alkaloid from corydalis extract | |
| CN104645667A (en) | Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof | |
| CN107688072B (en) | Detection method of Xingnaojing injection | |
| CN1321995C (en) | Total coptis alkaloid preparation method | |
| CN103961902A (en) | Simulated moving bed chromatographic separation system and method used for separating and concentrating target components from raw material liquid | |
| CN101791491A (en) | Online low-pressure, middle-pressure and high-pressure combined two-dimension preparation liquid chromatographic system | |
| CN104370895B (en) | A kind of preparation method of orientin and Lutonaretin | |
| CN112611825A (en) | Method and device for purifying plant polysaccharide by two-dimensional liquid chromatography | |
| CN109053433B (en) | Combined preparation method of ginkgolic acid | |
| CN109884199B (en) | Method for measuring content of flavonoid components in honey | |
| CN104784972A (en) | Preparation method and application of hesperidin immunoaffinity column | |
| CN205435056U (en) | Chromatographically pure piece -rate system | |
| CN101210039B (en) | Method for separating and preparing madecassoside chemical reference substance | |
| CN100526329C (en) | Production of high-purity peiminine and fritimine | |
| CN110066306A (en) | A kind of Isorhamnetin-3-O-neohespeidoside preparative liquid chromatography separation method | |
| CN100434135C (en) | Method for separating and purifying natural product using three-phase counter current chromatograph | |
| CN104193793A (en) | Method for preparing ginsenoside Rb3 in American ginseng fruit by adopting simulated moving bed | |
| CN114814034B (en) | Liquid chromatography method for simultaneously detecting contents of saponin and flavone in ophiopogon japonicus | |
| CN100422187C (en) | New method for extracting cephalotaxine and percephalotaxine | |
| CN103755674B (en) | The collaborative HSCCC of amination silica gel extracts the method for soybean isoflavone in soy sauce residues |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170208 Termination date: 20211118 |